Colicins

1
Colicins

• Yara Reis
• Erik Sommer
• Pascal Krämer
• Andreas Kühne
• Kathrin Nußbaum
• Marika Ziesack

2
Prey-Strain

• Double-transformation
• Sender for AI-1 and Sender for AI-2
• ? into DH5-a and Top 10
• Check via GFP-Measurement

3
Cloning Strategy for Killer

• 1. Cloning of the receiver-part into the
  pSB1A3-Vector
• - receiver-part via PCR from T9002
• - will have a Receiver-Vector

4
Cloning Strategy for Killer

• 2. Cloning of the Colicin-cassette behind the
  receiver-part into Receiver-Vector
• ? restartet cloning this week, that is why still
  in
• progress

Rec
Col
5
Test of colicins effect

• 2. Try
• Mixing Top10 cell suspension with colicin
  containing supernatant
• - Measurement of cell growth curves

6
Test of colicins effect

• 2. Try
• Results
• - no significant difference between normal
• growthcurve and that with colicin
  supernatant
• - perhaps the cells died because of mytomycin
  
• instead of colicin
• Advances
• - try other methods to stress colicin producing
  
• cells
• - measure decreasing GFP-expression of cells
• that shall be killed by colicin instead of
  OD

7
Future

• Cloning of colicin behind receiver
• ? verify and visualize
• Test of colicin effect on e.coli strains and
  aditionally on different eukaryotic cell lines
• Purify colicin-proteins via His-Tag for modeling
  parameters